One of the major problems in preserving organs for transplantation is maintaining cell viability during the period of time the organ is removed from the blood supply. In order to understand the events which accompany cell death during this period of time, the molecular mechanisms must be understood. The objectives of this study are to investigate the changes that occur in the concentration of all the nucleotides and nucleotide metabolism products which accompany an interruption in the oxygen supply to an organ. We will use recently developed methods in high pressure liquid chromatography to examine the effects of tissue injury on several aspects of nucleotide mtabolism. A decrease in nucleotide levels would affect many areas of cell function, and would play a major role in survival of the cell. One of the areas affected would be the transcription of the genetic material contained in the DNA. For faithful transcription of the DNA, all four nucleotides must be present in adequate concentrations as well as an active RNA polymerase and chromatin template. We have shown that RNA polymerase is very sensitive to ischemic injury; short periods of ischemia decrease activity of this enzyme dramatically. We will measure the effect of various storage solutions on RNA polymerase activity and will continue our investigation on the effect of temperature on RNA polymerase initiation and its importance to nuclear function and cell viability.